Simple Behavioral Assessment of Mouse Olfaction

Mu Yang1, Jacqueline N. Crawley1

1 National Institute of Mental Health, NIH, Bethesda, Maryland
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 8.24
DOI:  10.1002/0471142301.ns0824s48
Online Posting Date:  July, 2009
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This unit presents two basic protocols that offer rapid assessments of anosmia (the absence of a sense of smell) in mice. The buried food test is used to check for the ability to smell volatile odors. The olfactory habituation/dishabituation test is used to test whether the animal can detect and differentiate different odors, including both nonsocial and social odors. A non‐contact method of odor presentation, along with a general method for collecting urine samples, is given as an alternate protocol. The tests described in this unit only require simple equipment and can be adopted readily by most laboratories.Curr. Protoc. Neurosci. 48:8.24.1‐8.24.12. © 2009 by John Wiley & Sons, Inc.

Keywords: olfaction; mouse; anosmia; buried food test; habituation/dishabituation test

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Table of Contents

  • Introduction
  • Basic Protocol 1: Buried Food Test
  • Basic Protocol 2: The Olfactory Habituation/Dishabituation Test
  • Alternate Protocol 1: Testing Olfactory Habituation/Dishabituation with Noncontactable Odor Stimuli
  • Support Protocol 1: Urine Collection from Mice
  • Commentary
  • Literature Cited
  • Figures
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Basic Protocol 1: Buried Food Test

  • Mice: at least 6 weeks of age (juveniles may respond differently to food deprivation)
  • Food stimulus (particles should each weigh ∼1.5 g): Teddy Grahams (Nabisco) or other mini‐cookies of uniform size; other choices include sweetened cereal such as Froot Loops (Kellogg's), chocolate chips, squares of hard cheese, or small pellets of standard chow
  • Clean mouse cages (standard plastic cages, 46 cm L × 23.5 cm W × 20 cm H, or similar); the stainless steel top which holds food and water interferes with observation and should not be included in the setup.
  • Fresh cage bedding to create a 3‐cm layer in each cage
  • Clean holding cage for each subject for holding subject temporarily while stimulus is being buried; holding cages should be placed close to the testing cage
  • Digital timer: to monitor the test length (maximum = 15 min)
  • Traceable 300‐Memory Stopwatch (Control Company;

Basic Protocol 2: The Olfactory Habituation/Dishabituation Test

  • Mice: 6 weeks of age or older (sexually mature)
  • Odor stimuli:
    • Non‐social odors (use distilled H 2O as solvent): (1) distilled H 2O, (2) almond extract (McCormick; 1:100 dilution), (3) imitation banana extract (McCormick; 1:100 dilution), or similar; prepare fresh on day of experiment (almond and banana extract are selected because they are distinct yet neutral odors; to ensure consistent odor potency, replace concentrated extract every few months; store extracts away from direct sunlight and heat)
    • Social odors are obtained from mice of the same or opposite sex as the subject, depending on the scientific question (see below)
  • Clean mouse cages (standard plastic cages, 46 cm L × 23.5 cm W × 20 cm H, or similar), containing only fresh wood chip bedding
  • Clean plastic cage lids, one for each cage
  • Cotton‐tipped wooden applicators (6 in. length; Solon Manufacturing Company,, or similar
  • Small (4.5 × 4.5 cm) weigh boat
  • Digital stopwatches (silenced, as described in protocol 1) to record sniffing time
  • Digital timer to register the 2‐min trial duration
  • Glass vials for water and solutions (label clearly and keep tightly capped when not in use)
  • Large lidded jar for disposal of used applicators (a temporary disposal container helps to prevent cross‐trial contamination; empty jar after testing each animal)

Alternate Protocol 1: Testing Olfactory Habituation/Dishabituation with Noncontactable Odor Stimuli

  • Mice: at least 6 weeks of age (sexually mature)
  • Odorant (e.g., urine, see protocol 4)
  • Fine wire mesh
  • Clean mouse cages (standard plastic cages, 46 cm L × 23.5 cm W × 20 cm H, or similar), containing only fresh wood chip bedding
  • Cage lids, including the plastic top and the stainless steel grid top
  • Small plastic weigh boats (4.5 cm × 4.5 cm)
  • Filter paper, cut into individual squares (2 cm × 2 cm)
  • Double‐sided tape
  • Digital stopwatches (silenced, as described in protocol 1)
  • Digital timer to register trial duration
  • Large lidded jar to dispose of pipet tips and pieces of filter paper

Support Protocol 1: Urine Collection from Mice

  • Adult mice, 6 weeks of age or older
  • Microcentrifuge tubes or clean glass vials
  • Whatman no. 5 filter paper
  • Small sealable plastic bags
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Literature Cited

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